LOCUS       V044455                 9191 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044455
VERSION     V044455
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..3552
                     /codon_start=1
                     /gene=""
                     /note=""
                     /product="human lysosome-associated membrane glycoprotein
                     1"
                     /protein_id=""
                     /transl_table=1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASA,AMFMVKNGNGTACIMA
                     NFSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLN
                     FTRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVH
                     MNNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDK
                     YNVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHS
                     EGTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCN
                     AEEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLI
                     VLIAYLVGRKRSHAGYQTI"
                     /label="LAMP1"
     CDS             3595..4305
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced monomeric blue fluorescent protein
                     (Subach et al., 2011)"
                     /transl_table=1
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
                     /label="mTagBFP2"
     promoter        4356..4567
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4580..4848
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4881..5279
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5286..5874
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5948..6181
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6253..6374
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6393..6528
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6566..6582)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6590..6606
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6614..6644)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6659..6680
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6968..7556)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7727..8587)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8588..8692)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9166..9182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggcggcc cccggcagcg cccggcgacc cctgctgctg
     2341 ctactgctgt tgctgctgct cggcctcatg cattgtgcgt cagcagcaat gtttatggtg
     2401 aaaaatggca acgggaccgc gtgcataatg gccaacttct ctgctgcctt ctcagtgaac
     2461 tacgacacca agagtggccc taagaacatg acctttgacc tgccatcaga tgccacagtg
     2521 gtgctcaacc gcagctcctg tggaaaagag aacacttctg accccagtct cgtgattgct
     2581 tttggaagag gacatacact cactctcaat ttcacgagaa atgcaacacg ttacagcgtc
     2641 cagctcatga gttttgttta taacttgtca gacacacacc ttttccccaa tgcgagctcc
     2701 aaagaaatca agactgtgga atctataact gacatcaggg cagatataga taaaaaatac
     2761 agatgtgtta gtggcaccca ggtccacatg aacaacgtga ccgtaacgct ccatgatgcc
     2821 accatccagg cgtacctttc caacagcagc ttcagccggg gagagacacg ctgtgaacaa
     2881 gacaggcctt ccccaaccac agcgccccct gcgccaccca gcccctcgcc ctcacccgtg
     2941 cccaagagcc cctctgtgga caagtacaac gtgagcggca ccaacgggac ctgcctgctg
     3001 gccagcatgg ggctgcagct gaacctcacc tatgagagga aggacaacac gacggtgaca
     3061 aggcttctca acatcaaccc caacaagacc tcggccagcg ggagctgcgg cgcccacctg
     3121 gtgactctgg agctgcacag cgagggcacc accgtcctgc tcttccagtt cgggatgaat
     3181 gcaagttcta gccggttttt cctacaagga atccagttga atacaattct tcctgacgcc
     3241 agagaccctg cctttaaagc tgccaacggc tccctgcgag cgctgcaggc cacagtcggc
     3301 aattcctaca agtgcaacgc ggaggagcac gtccgtgtca cgaaggcgtt ttcagtcaat
     3361 atattcaaag tgtgggtcca ggctttcaag gtggaaggtg gccagtttgg ctctgtggag
     3421 gagtgtctgc tggacgagaa cagcatgctg atccccatcg ctgtgggtgg tgccctggcg
     3481 gggctggtcc tcatcgtcct catcgcctac ctcgtcggca ggaagaggag tcacgcaggc
     3541 taccagacta tcggatccgg cggtggagga agcggcggag gtggatctct cgagatggtg
     3601 tctaagggcg aagagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     3661 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     3721 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     3781 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
     3841 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
     3901 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     3961 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
     4021 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
     4081 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacgccaa gaccacatat
     4141 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
     4201 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     4261 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattaagg atccgcggcc
     4321 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     4381 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     4441 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     4501 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     4561 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     4621 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     4681 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     4741 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     4801 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc
     4861 ggcgcctacg ctaggccacc atggccaagc ctttgtctca agaagaatcc accctcattg
     4921 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
     4981 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     5041 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     5101 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     5161 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     5221 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag
     5281 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5341 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5401 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5461 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5521 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5581 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5641 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     5701 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5761 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5821 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta
     5881 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     5941 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact
     6001 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6061 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6181 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6241 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6301 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6361 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     6421 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6481 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     6541 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6601 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6661 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6721 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6781 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6841 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6901 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6961 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7021 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7081 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7141 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7381 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7741 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7801 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7861 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7921 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7981 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8041 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8101 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8161 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8221 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8281 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8341 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8401 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8461 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8521 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8581 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8641 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8701 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8761 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     8821 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     8881 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     8941 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     9001 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     9061 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     9121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     9181 gtgccaagct g
//
